1. Volume 22, Issue 2, Pages 464-475 (February 2014)
Vaccine-elicited Human T Cells Recognizing Conserved Protein Regions Inhibit HIV-1 
Nicola Borthwick, Tina Ahmed, Beatrice Ondondo, Peter Hayes, Annie Rose, Umar Ebrahimsa, Emma-Jo Hayton, Antony Black, Anne Bridgeman, Maximillian Rosario, Adrian VS Hill, Eleanor Berrie, Sarah Moyle, Nicole Frahm, Josephine Cox, Stefano Colloca, Alfredo Nicosia, Jill Gilmour, Andrew J McMichael, Lucy Dorrell, Tomáš Hanke 
Molecular Therapy 
Volume 22, Issue 2, Pages (February 2014)
DOI: /mt
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions

2. Figure 1
The HIVconsv immunogen and trial design. (a) A schematic representation of the conserved region immunogen HIVconsv. Capital letters above bars indicate the clade used for the derivation of the consensus aa sequence for each conserved segment.16 Below are the regions of the HIVconsv protein covered by overlapping 15/11 peptide pools used in enzyme linked immunoabsorbent spot (ELISPOT) and intracellular cytokine staining (ICS) immunological assays. HIV-1 protein origin of conserved regions are color-coded with navy blue indicating Mamu-A*01 and H-2Dd–restricted T-cell epitopes, and mAb Pk tag added to facilitate protein detection. (b) Trial design. Four immunization schedules used in the HIV-CORE 002 clinical trial are shown, whereby c and C are low and regular doses of ChAdV63.HIVconsv, respectively, D is pSG2.HIVconsv DNA, M is MVA.HIVconsv and P is placebo. The vaccine-to-placebo ratio was 4:1. Note that a longer gap of 8 weeks was used after the ChAdV63.HIVconsv administration before MVA.HIVconsv boost.50
Molecular Therapy  , DOI: ( /mt )
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions

3. Figure 2
Vaccine-elicited frequencies of T cells recognizing conserved regions of HIV-1. Freshly isolated, unexpanded peripheral blood mononuclear cells (PBMC) were stimulated with 6 pools of overlapping 15/11 peptide pools derived from the HIVconsv immunogen (Figure 1a) in an IFN-γ enzyme linked immunoabsorbent spot (ELISPOT) assay and the sums of net peptide Pool 1–6 frequencies are plotted for individual volunteers for each time point. S, screening visit maximum 2 weeks before vaccination. Timing of vaccine administrations is indicated below. Volunteers are color-coded and boxed numbers identify placebo recipients. c and C are low and regular doses of ChAdV63.HIVconsv, respectively; D is pSG2.HIVconsv DNA; and M is MVA.HIVconsv.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions

4. Figure 3
The breadth of vaccine-elicited T cells. Freshly isolated, unexpanded peripheral blood mononuclear cells (PBMC) were stimulated with Pools 1–6 of overlapping 15/11 peptides derived from the HIVconsv immunogen (Figure 1a) or the FEC (influenza virus, EBV, CMV) pool (color-coded) in an IFN-γ enzyme linked immunoabsorbent spot (ELISPOT) assay and the net frequencies of cells specific for each pool are shown. S is screening visit. Volunteer numbers are shown above the graphs and boxed numbers indicate placebo recipients.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions

5. Figure 4
Breadth of vaccine-elicited responses in groups CM and DDDCM. Stimulatory peptides were identified using a cultured IFN-γ enzyme linked immunoabsorbent spot (ELISPOT) assay and are shown as a heat map for individual vaccines. Frozen peripheral blood mononuclear cells (PBMC) from week time points were cultured for 10 days using HIVconsv Pools 1–6 and assayed using individual peptides. Numbers in each box show SFU/106 of cultured cells. In the first column, peptide numbers are color-coded as for their origin: blue, Gag; pink, Pol; yellow, Vif; purple, Env. In the first and second columns, peptides containing a junction indication by a hyphen are shown on orange background. Black horizontal lines separate Pools 1–6. For volunteer no. 405, not enough cells were available for epitope mapping. Volunteers’ HLA class I types are shown on the top of the figure.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions

6. Figure 5
In vitro HIV-1 inhibition by vaccine-elicited CD8+ T cells. (a) HIV-1 inhibition by two individuals with broad VIA activity. Eight available viruses employed in this assay are listed and color-coded on the right. Dotted lines show the highest background inhibition (1.3 log10). (b) HIV-1 inhibition by individual volunteers for the two most commonly inhibited viruses indicated above the graphs. Dotted lines show the highest background inhibition for each virus of all tested individuals. Boxed volunteer numbers indicate placebo recipients.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions

7. Figure 6
Replicative capacity of HIV-1 isolates in autologous CD4+ cells. HIV-1 growth in expanded autologous CD4+ cells was measured by the p24 concentration in the tissue culture supernatant on day 13. Broad controllers numbers 411 and 418 are highlighted in red and green, respectively. Blue bars indicate the average concentration for individual viruses.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions

8. Figure 7
Correlation of frequencies of Gag- and Pol-specific CD8+ effector cells with virus inhibition. (a) Peripheral blood mononuclear cells (PBMC) from pre- and three postvaccination time points were nonspecifically expanded by incubation with anti-CD3/CD4 bi-specific antibody and subjected to an IFN-γ enzyme linked immunoabsorbent spot (ELISPOT) assay using 4 pools of overlapping 15/11 peptides derived from Gag, 2 pools of Pol and Env+Vif. (The same cultured cells were used for virus-inhibition assay (VIA) shown in Figure 5.) Individual volunteers’ frequencies are color-coded with boxed volunteer numbers indicating placebo recipients. (b) For the two most commonly inhibited viruses indicated above the graphs, correlations of nonspecifically expanded CD8+ effector frequencies specific for Gag, Pol, and Env+Vif with overall, i.e., by cells specific for Gag, Pol, Env, and Vif together, virus in vitro inhibition are shown for the peak inhibition time points. Red dots, CM; purple squares, DDDCM; blue triangles, DDDMC. For each correlation, Spearman rank (r) and P (two-tailed) values are shown.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2014 The American Society of Gene & Cell Therapy Terms and Conditions